pET15b-pep-1-p27mt的构建、表达及意义  

Construction and Expression of pET15b-pep-1-p27mt Recombinant

作　　者：王立林[1] 严世荣[2] 汪炳华[1] 王家宁[2] 黄永章[2] 

机构地区：[1]武汉大学医学院,湖北武汉430000 [2]郧阳医学院附属人民医院,湖北十堰442000

出　　处：《药物生物技术》2005年第5期285-290,共6页Pharmaceutical Biotechnology

基　　金：湖北省教育厅资助(编号:Q200524001)

摘　　要：构建细胞穿膜肽(cell-penetrating peptides,CPPs)PEP-1和细胞周期抑制蛋白p27mt的重组体,并在大肠杆菌中表达。将已构建的表达细胞周期抑制蛋白p27mt的基因,克隆入pET15b-pep-1原核表达载体,在大肠杆菌BL21(DE3)LysS内,以IPTG诱导融合蛋白表达。表达产物用SDS-PAGE及Western blot分析鉴定。结果:成功地构建PEP-1-p27mt融合蛋白原核表达载体,并在IPTG诱导下获得特异性的表达。PEP-1-p27mt融合蛋白表达载体的构建,为体内应用细胞周期抑制蛋白诱导肿瘤细胞凋亡提供了理论基础。The aim is to construct prokaryotic expression of pET15b-pep-1-p27mt recombinant and to express it in E. coli. The cDNAs encoding cyclin dependent kinase inhibitors p27mt was cloned into prokaryotic expression vector pET15b. Recombinant plasmid was transformed into E. col i BL21 （DE3）LysS, then was induced with IPTG. The expression of the fusion protein was analyzed by SDS-PAGE and Western blot. The recombinant plasmids were constructed and expressed after IPTG inducting successfully. The obtained PEP-1-p27mt fusion protein has laid the foundation for using cyclin dependent kinase inhibitors p27mt in the inducing apoptosis of tumor cells.

关 键 词：细胞穿膜肽 PEP-1 PEP-1-p27mt融合蛋白 pET15b-pep-1-p27mt 重组表达子 抑癌基因 

分 类 号：Q786[生物学—分子生物学]